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利用针对小鼠H-2 I类抗原预测羧基末端的抗体检测其分泌形式。

Detection of a secreted form of the murine H-2 class I antigen with an antibody against its predicted carboxyl terminus.

作者信息

Maloy W L, Coligan J E, Barra Y, Jay G

出版信息

Proc Natl Acad Sci U S A. 1984 Feb;81(4):1216-20. doi: 10.1073/pnas.81.4.1216.

DOI:10.1073/pnas.81.4.1216
PMID:6583704
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC344797/
Abstract

Analysis of H-2 class I-specific cDNA clones has suggested the synthesis by the liver of a class I molecule that is secreted rather than membrane bound. To detect this putative class I-related molecule, we have predicted a unique region of amino acid sequence located toward the carboxyl terminus of the molecule that is not expected to be shared with any of the classical H-2 class I antigens, and we have generated specific antibodies to a synthetic peptide corresponding to this sequence. Indirect immunoprecipitation with this antibody led to the identification of a Mr 40,000 polypeptide in association with beta 2-microglobulin in the serum of mice of five different H-2 haplotypes. This class I molecule is also detected in the liver together with lower molecular weight components, which are presumably underglycosylated precursors. Synthesis of this molecule is not detected in thymus, spleen, kidney, or testis. This class I serum component has no detectable reactivity with either a broad-specificity alloantiserum against H-2b or a xenoantiserum against purified H-2a class I molecules. The availability of a specific antibody against the secreted class I molecule offers a means to purify this protein for structural and functional studies.

摘要

对H-2 I类特异性cDNA克隆的分析表明，肝脏合成了一种分泌型而非膜结合型的I类分子。为了检测这种假定的与I类相关的分子，我们预测了该分子羧基末端的一个独特氨基酸序列区域，预计该区域不会与任何经典的H-2 I类抗原共有，并且我们针对与该序列对应的合成肽产生了特异性抗体。用该抗体进行间接免疫沉淀，在五种不同H-2单倍型小鼠的血清中鉴定出一种与β2-微球蛋白相关的40,000道尔顿的多肽。在肝脏中也检测到了这种I类分子以及较低分子量的成分，这些成分可能是低糖基化的前体。在胸腺、脾脏、肾脏或睾丸中未检测到该分子的合成。这种I类血清成分与针对H-2b的广泛特异性同种异体抗血清或针对纯化的H-2a I类分子的异种抗血清均无可检测的反应性。针对分泌型I类分子的特异性抗体的可用性为纯化该蛋白质以进行结构和功能研究提供了一种方法。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/716b/344797/4ed627ad8ea0/pnas00605-0243-b.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/716b/344797/52d06221ab51/pnas00605-0241-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/716b/344797/7eaf6d155805/pnas00605-0242-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/716b/344797/b2a117e30eae/pnas00605-0242-b.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/716b/344797/154af7c03d57/pnas00605-0243-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/716b/344797/4ed627ad8ea0/pnas00605-0243-b.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/716b/344797/52d06221ab51/pnas00605-0241-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/716b/344797/7eaf6d155805/pnas00605-0242-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/716b/344797/b2a117e30eae/pnas00605-0242-b.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/716b/344797/154af7c03d57/pnas00605-0243-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/716b/344797/4ed627ad8ea0/pnas00605-0243-b.jpg

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